• 한국가축번식학회지
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• 제17권4호
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• pp.331-337
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• 1994

처녀발생은 난자의 세포질성숙을 투명대경화나 체외수정에 있어 난자 외적요소의 문제점을 배제하고 측정할 수 있는 지표이다. 본 실험에서는 체외성숙시간에 따른 소 난자의 처녀발생활성을 조사하였다. 도살장 난소로부터 회수한 미성숙난포란을 15% 소 태아혈청이 첨가된 TCM 199에서 6시간 간격으로 24~48시간까지 성숙시킨 후 7% ethanol로 7분간 활성화시켰다. 핵성숙과 세포질성숙은 rapid staining에 의해 핵형태와 전핵의 형성 유무로 판정하였다 핵성숙율은 24~48시간 사이 각각 81, 89, 72, 60 및 60%로 체외성숙 36시간에 성숙율이 최고였으나, 반면 감수분열 중기 II 염색체이상은 36시간부터 증가(0~30%)하였다. 에탄올처리에 의한 전핵형성율은 체외성숙 24~48시간에 각각 67, 68, 73, 84 및 87%였고, 그 중 이배체율은 각각 4, 5, 10, 16 및 20%로 성숙시간이 증가함에 따라 증가하였다. 위 실험의 결과 난자의 체외성숙 연장에 따라 전핵형성과 이배체수가 증가되는 것으로 나타났으며, 정상적인 핵성숙에 비해 세포질성숙은 더 많은 성숙시간이 필요한 것으로 나타났다. 이러한 결과들은 소 초기배 체외생산시와 핵치환용 핵수용란 생산시 적정 성숙시간 결정에 유용하게 이용될 수 있을 것이다.

• 한국가축번식학회지
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• 제16권1호
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• pp.7-14
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• 1992

These experiments were designed to investigate the effects of glycosaminoglycans(GAGs), which maybe produced by cumulus cells, on in vitro maturation process of bovine follicular oocytes. The rates of in vitro maturation of bovine oocytes were examined after incubating with the various concentrations of a hyaluronic acid, chondroitin sulfate, or heparin for 26 hours. The results obatained in these experiments were summarized as follows : 1. When the cumulus or removed oocytes treated with hyaluronic acid(200, 400, 800 and 1, 600$\mu\textrm{g}$/ml), the maturation rates of cumulus-removed oocytes were 48.6, 59.4, 68.4 and 61.3% respectively. Especially, at a concentration of 800$\mu\textrm{g}$/ml, the maturation rates(68.8%) of cumulus-removed oocytes were slightly lower than that(78.3%) of cumulus-enclosed oocytes. However, the treatment of hyaluronic acid showed significantly higher maturation rates compared to that(48.4%) of control group of cumulus-removed oocytes(p<0.05). Therefore, these results suggest thate hyaluronic acid had a beneficial effect on maturation of bovine follicular oocytes. 2. The maturation rate of cumulus-free oocytes treated with chondroitin sulfate(200, 400, 800 and 1, 600$\mu\textrm{g}$/ml) was 52.3, 56.1, 55.9 and 52.2% respectively. These rates were not different from those(52.9%) of control group. However, these rates in cumulus-free oocytes were significantly lower than that(79.0%) in cumulus-enclosed oocytes(p<0.01). Therefore, these results suggest that chondroitin sulfate do not affect on the maturation of oocytes. 3. In cumulus-free oocytes cultured with different heparin concentration, the maturation rates were ranged from 48.5 to 52.1%, showing no differencies from that(50.7%) of control group. However, these rates were significantly lower than 80.0% in cumulus-enlosed oocytes(p<0.01). 4. The nuclear maturaton of oocytes was increased by treatment of each of three glycosaminoglycans(hyaluronic acid, chondroitin sulfate, and heparin). In addition, the treatment of mixed together showed the significant additive effect. Hyaluronic acid was more effective than chondroitin sulfate and heparin were.

• 한국동물생명공학회지
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• 제36권3호
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• pp.162-167
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• 2021

Camel (camelus dromedarius) is a unique large mammalian species that can survive harsh environmental conditions and produce milk, meat, and wool. Camel reproduction is inferior when compared to other farm animal species such as cattle and sheep. Several trials have been reported to increase camel reproduction and production through assisted reproductive techniques (ARTs) such as in vitro fertilization and cloning. For these reasons, obtaining enough mature oocytes is a cornerstone for ARTs. This demand would be improved by the oocyte in vitro maturation (IVM) systems. In this review, the current approaches and views from different laboratories using ARTs and the IVM to produce embryos in vitro in camel species. For the last two decades, conventional IVM system was the common approach, however, recently the bi-phasic IVM system has been introduced and showed promising improvement in IVM of camel oocytes. Detailed studies are needed to understand camel meiosis and IVM to efficiently increase the production of this species.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.242-242
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• 2004

This study was conducted to investigate the effects of various supplements added into maturation medium of immature porcine oocytes on quantity of cytoplasmic lipid droplets(LD), subsequent fertilization and development to the blastocyst stage in vitro. The basic maturation medium was TCM 199 + 1 ㎍/㎖ FSH, 0.57 mM cystein, 10 ng/㎖ EGF and was supplemented various supplements(10% FBS, 10% pFF, 0.4% BSA, 1.0% BSA, 0.4% PVP, 1.0% PVP). (omitted)

• 한국수정란이식학회지
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• 제32권3호
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• pp.139-146
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• 2017

The objective of this study was to determine the effect of fructose that was supplemented to a chemically defined in vitro maturation (IVM) medium on oocyte maturation and embryonic development after parthenogenesis in pigs. The base medium for in vitro maturation (IVM) was porcine zygote medium (PZM) that was supplemented with 0.05% (w/v) polyvinyl alcohol (PVA) or 10% (v/v) porcine follicular fluid (pFF). In the first experiment, when immature pig oocytes were matured in a chemically defined medium that was supplemented with 5.5 mM glucose or with 1.5, 3.0 and 5.5 mM fructose, 3.0 mM fructose resulted in a higher nuclear maturation (91.5%) than 1.5 and 5.5 mM fructose (81.9 and 81.9%, respectively) but showed a similar result with 5.5 mM glucose (94.2%). However, there was no significant differences among groups in the embryo cleavage (89.4-92.4%), blastocyst formation (37.5-41.1%), and mean cell number of blastocyst (30.8-34.2 cells). Fructose at the concentration of 3.0 mM (1.08 pixels/oocyte) resulted in a higher intra-oocyte glutathione (GSH) content than 1.5 and 5.5 mM fructose (1.00 and 0.87 pixels/oocytes, respectively) while the cumulus cell expansion was not influenced. In the second experiment, effect of individual and combined supplementation of a chemically defined maturation medium with 5.5 mM glucose or 3.0 mM fructose was examined. No significant effect was found in the nuclear maturation (86.3-92.6%). Embryo cleavage was significantly increased by the combined supplementation with glucose and fructose (95.2%) compared to that with 3.0 mM fructose only (85.7%) while blastocyst formation (37.3-42.8%) and embryonic cell number (33.3-34.1 cells) were not altered. Effect of supplementation of pFF-containing medium with glucose and fructose + glucose was examined in the third experiment. No significant effect by the supplementation with glucose and fructose or glucose alone was observed in the nuclear maturation of oocytes (90.7-94.1%) and blastocyst formation (51.0-56.5%). Our results demonstrate that 3.0 mM fructose was comparable to 5.5 mM glucose in supporting in vitro oocyte maturation and embryonic development after parthenogenesis and could be used as an alternative energy source to glucose for in vitro maturation of pig oocytes.

• Reproductive and Developmental Biology
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• 제33권3호
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• pp.125-131
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• 2009

The aim of this study is to investigate the effect of porcine epididymal fluid (pEF) on in vitro-maturation and subsequent fertilization of porcine follicular oocytes. Porcine cumulus-oocytes complexes retrieved from antral follicles were cultured in tissue culture medium (TCM)-l99 supplemented with pEF of different concentrations. At 48 h after culture, development of oocytes to germinal vesicle (GV) breakdown, metaphase I, anaphase-telophase I, and metaphase II were examined Significant (p<0.05) increase in the proportion of oocytes developed to MII stage was observed in oocytes cultured in pEF-containing TCM-l99 than in oocytes cultured in pEF-free TCM-199 (46.2% vs 16.7%), which was a dose-dependent manner. Subsequently, the proportion of monospermic fertilization were significantly (p<0.05) increased in oocytes cultured in the TCM supplemented with pEF than those cultured in pEF-free TCM-199 (51.0% vs 24.1%). In the second series of experiment, the percentage of MII oocytes was significantly (p<0.05) increased after exposure of oocytes to pEF during the first 22 h period of culture than after exposure of oocytes to pEF during the next 24 h of culture, while no significant difference in the percentage of monospermy was observed. The results of this study demonstrate that pEF contains at least enhancing component(s) for nuclear maturation.

• 농업과학연구
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• 제44권3호
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• pp.309-317
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• 2017

Melatonin (N-acetyl-5-methoxytryptamine) is an indole synthesized from tryptophan by the pineal gland in animal. The major function of melatonin is to modulate circadian and circannual rhythms in photoperiodic mammals. Importantly, however, melatonin is also a free radical scavenger, anti-oxidant, and anti-apoptotic agent. Recently, the beneficial effects of melatonin on oocyte maturation and embryonic development in vitro have been reported in many species such as pig, cattle, sheep, mouse, and human. In this review, we will discuss recent studies about the role of melatonin in the production of porcine and bovine oocytes and embryos in vitro in order to provide useful information of melatonin in oocyte maturation and embryo culture in vitro.

• 한국수정란이식학회지
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• 제18권1호
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• pp.75-80
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• 2003

• 한국수정란이식학회지
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• 제15권1호
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• pp.39-46
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• 2000

This study was conducted to investigate the effect of hormones, protein sources and anti-oxidants on in vitro maturation (IVM) and in vitro fertilization(IVF) of bovine follicular oocytes. The rates of Holstein follicular oocytes classified as grade A and B(50.2% and 33.2%) were higher than those of Hanwoo cattle(40.3% and 32.0%, P<0.05). The cumulus cell expansion rates of oocytes cultured in TCM-199 and Ham's F-10 medium supplemented with 10% FCS and hormones were higher (81.9~87.6%) than those of non-treated groups (74.5~81.7%). The fertilization rates of oocytes cultured in TCM-199 and Ham's F-10 medim supplemented with 10% FCS, 1% BSA and 10% bFF was 53.8~55.0%, 51.4~52.6%, and 47.0~50.0%, respectively. The polyspermy rates was 13.6~14.2%, 10.0~11.1%, and 10.0%, respectively. When the oocytes were cultured in TCM-199 and Ham's F-10 medium with 50${\mu}{\textrm}{m}$ $\alpha$-tocopherol, the fertilization rates was 62.0 and 60.2%, respectively. In the maturation medium added of 100${\mu}{\textrm}{m}$ cysteamine, the fertilization rates was 64.7 and 66.7%, respectively. The fertilization and polyspermy rates of treated groups were higher than those of non-treated group. The results show that hormones, protein sources and anti-oxidants can provide a benefit for in vitro maturation and fertilization of bovine follicular oocytes.

• 한국발생생물학회지:발생과생식
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• 제12권2호
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• pp.125-132
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• 2008

멜라토닌(N-acetyl-5-methoxytryptamine)은 포유동물의 뇌의 송과선에서 분비되는 호르몬으로 수면과 생체 리듬 등을 조절하고 난소 기능과 번식에도 영향을 미친다. 또한, 강력한 scavenger로서 항산화제의 역할을 한다. 이 연구의 목적은 멜라토닌이 생쥐 난구세포-핵낭(germinal vesicle, GV) 시기 난자 복합체의 체외성숙에 미치는 영향을 알아보는 것이다. 3주령의 ICR 암컷 생쥐의 난소에서 회수된 난자-난구세포 복합체를 0, 0.1 nM, 10 nM, 1,000 nM의 멜라토닌이 첨가된 배양액에서 18시간 동안 배양하고, 제1극체의 방출 여부를 확인하여 성숙율을 확인하였다. 체외성숙 후 TUNEL assay와 성숙율이 가장 높게 나타났다. 체외성숙된 난자에서는 세포자연사가 나타나지 않았으나 난구세포에서는 관찰되었으며, 1,000 nM을 첨가하여 배양한 군의 난구세포는 유의하게 낮은 세포자연사를 나타냈다. 그리고 1,000 nM의 멜라토닌을 첨가한 군의 난구세포에서 멜라토닌 수용체의 mRNA가 대조군에 비해 낮게 발현되었다. 이상의 결과를 종합하면 체외성숙 배양액에 첨가된 멜라토닌은 난구세포의 세포자연사를 줄여줌으로써 생쥐 미성숙 난자의 체외성숙을 향상시키는 역할을 하는 것으로 사료된다.